Figure 1. Multiplex qRT-PCR using the RealHelix™ qRT-PCR Kit [v6a] [UDG System].

10-fold dilutions of purified SARS-CoV-2 viral RNA (from 10^-4 to 10^-7) spiked into a swab sample were analyzed using three different sets of target-specific primers and probes (N gene, ORF8, and S gene). The amount of viral RNA used in the 10^-7 dilution fold corresponds to approximately 10² copies. The probes were labeled with Texas Red (N gene), JOE (ORF8), FAM (S gene). The multiplex reaction was tested using the RealHelix™ qRT-PCR Kit [v6a] [UDG System] on a real-time PCR instrument, Bio-Rad CFX96.

Figure 2. Multiplex qRT-PCR using the RealHelix™ qRT-PCR Kit [v6a] [UDG System].

Plasmid clone mixtures harboring the partial region for each SARS-CoV-2 were used as templates in this reaction. SARS-CoV-2 clone mixture (10⁶, 10⁵, 10⁴, 10³, 10² copies) were analyzed using three different sets of target-specific primers and probes (N gene, ORF8 and S gene). The probes were labeled with Texas Red (N gene), JOE (ORF8), and FAM (S gene). The multiplex reaction was tested on a real-time PCR instrument, Bio-Rad CFX96.